Enabling immunotherapy for high-risk Group 3 medulloblastoma via systems immunology

通过系统免疫学对高危 3 组髓母细胞瘤进行免疫治疗

• 批准号: 10714138
• 负责人: Hongbo Chi
• 金额: $ 81.9万
• 依托单位: ST. JUDE CHILDREN'S RESEARCH HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-07 至 2028-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10714138
• 关键词: Address; Adoptive Cell Transfers; Atlases; Biology; Brain Neoplasms; CAR T cell therapy; CD8-Positive T-Lymphocytes; CRISPR screen; Cancer Etiology; Cell Communication; Cell Therapy; Cell physiology; Cells; Cellular biology; Child; Childhood Malignant Brain Tumor; Classification; Clustered Regularly Interspaced Short Palindromic Repeats; Data; Development; Disease; Exclusion; Exhibits; Genetic; Genetically Engineered Mouse; Genomics; Goals; Heterogeneity; Human; Immune; Immune Evasion; Immunocompetent; Immunology; Immunotherapeutic agent; Immunotherapy; Joints; Macrophage; Maps; Mediating; Modeling; Molecular; Multiomic Data; Mus; Network-based; PTPRC gene; Patients; Phenotype; Pre-Clinical Model; Proteomics; Refractory; Resistance; Reverse engineering; Saint Jude Children' s Research Hospital; Sampling; Signal Transduction; Sorting; Subgroup; System; Systems Biology; T cell response; T cell therapy; T-Lymphocyte; TCF Transcription Factor; Technology; Testing; Tumor Immunity; Tumor stage; Tumor-infiltrating immune cells; Visualization; cancer type; childhood cancer mortality; chimeric antigen receptor T cells; clinically relevant; cloud based; data visualization; disorder risk; experimental study; functional genomics; high risk; high risk population; improved; in vivo; insight; medulloblastoma; mouse model; multiple omics; neoplastic cell; nerve stem cell; notch protein; novel; overexpression; screening; single cell analysis; single-cell RNA sequencing; spatiotemporal; stem-like cell; tool; transcriptomic profiling; transcriptomics; tumor; tumor microenvironment; tumor progression; tumor-immune system interactions

PROJECT SUMMARY / ABSTRACT The goal of this project is to dissect the immune evasion mechanisms and enable immunotherapy for children with high-risk Group 3 medulloblastoma (G3MB) via systems immunology approaches. Brain tumors are the leading cause of cancer-related deaths in children. Medulloblastoma is the most prevalent malignant pediatric brain tumor and is characterized by four major molecular subgroups, among which G3MB is the most aggressive form and features MYC overexpression. The immunosuppressive tumor microenvironment (TME) is poorly understood in G3MB, and no immunotherapy is available for children with this high-risk disease. Systems immunology approaches—especially single-cell and spatial multi-omics profiling and in vivo CRISPR-based functional screening—have proven powerful in dissecting tumor–TME interactions and identifying novel immunotherapy targets in various cancer types, but very few studies have integrated these approaches. In our preliminary studies, we applied two unique immunocompetent genetically-engineered mouse models (GEMMs) of MYC-driven G3MB and performed scRNA-seq, scATAC-seq and spatial transcriptomics profiling. We enriched immune cells from the TME by sorting CD45 positive cells for single-cell studies. Our preliminary analysis of single-cell and spatial omics data revealed striking interactions of neural stem cell-like tumor cells with macrophages and other immune cells that potentially create a suppressive TME and drive immune evasion in mouse G3MB. We also performed in vivo CRISPR screening in tumor cells using the GEMMs to identify modulators of tumor development, which demonstrated the feasibility of in vivo functional genomics screening in our preclinical models. In this project, first, we propose to utilize cutting-edge single-cell and spatial omics technologies to characterize the two G3MB GEMMs at different stages of tumor progression. We will use our network-based tools to integrate these multi-omics data to dissect the dynamic tumor–immune interactions and underlying “hidden” drivers that drive the immune exclusion and suppression during G3MB progression. We will also validate discoveries of G3MB from mouse studies in patient samples. We will develop a cloud-based portal to visualize and explore our single-cell and spatial data and tumor–TME interactomes of G3MB. Second, we will establish the mechanistic basis of tumor–T cell interactions and strategies to enable adoptive T cell therapy for G3MB by discovering functional drivers and putative targets in both tumor cells and T cells. To this end, we will apply both candidate approach and in vivo CRISPR screening in immunocompetent GEMMs to identify tumor- intrinsic modulators that will remodel the suppressive TME and sensitize G3MB tumors to adoptive T cell and CAR-T cell therapies. We will also test if targeting inhibitory factors for T cell function will enable and optimize effective adoptive T cell therapies against such tumors. Our studies promise to provide new insights into mechanisms of tumor–TME interactions in G3MB and manifest legitimate immunotherapeutic opportunities.

项目摘要/摘要 该项目的目标是剖析免疫逃避机制，使儿童免疫治疗成为可能。 与高危组3髓母细胞瘤（G3 MB）通过系统免疫学方法。脑肿瘤是 儿童癌症相关死亡的主要原因。髓母细胞瘤是儿童最常见的恶性肿瘤 脑肿瘤，其特征在于四个主要的分子亚群，其中G3 MB是最具侵袭性的 形式和特征MYC过度表达。免疫抑制性肿瘤微环境（TME） 在G3 MB中了解，并且没有免疫疗法可用于患有这种高危疾病的儿童。系统 免疫学方法-特别是单细胞和空间多组学分析和基于CRISPR的体内 功能筛选-已被证明在解剖肿瘤-TME相互作用和鉴定新的 免疫疗法靶向于各种癌症类型，但很少有研究整合了这些方法。在我们 在初步研究中，我们应用了两种独特的免疫活性基因工程小鼠模型（GEMM） 的MYC驱动的G3 MB，并进行scRNA-seq，scATAC-seq和空间转录组学分析。我们丰富了 通过分选用于单细胞研究的CD 45阳性细胞从TME中分离免疫细胞。我们的初步分析 单细胞和空间组学数据揭示了神经干细胞样肿瘤细胞与 巨噬细胞和其他免疫细胞可能会产生抑制性TME并驱动免疫逃避， 小鼠G3 MB。我们还使用GEMM在肿瘤细胞中进行了体内CRISPR筛选，以识别 肿瘤发展的调节剂，这证明了体内功能基因组学筛选的可行性 在我们的临床前模型中。在这个项目中，首先，我们建议利用尖端的单细胞和空间组学 技术来表征肿瘤进展不同阶段的两种G3 MB GEMM。我们将用我们 基于网络的工具来整合这些多组学数据，以剖析动态肿瘤-免疫相互作用， 在G3 MB进展期间驱动免疫排斥和抑制的潜在“隐藏”驱动因素。我们将 还验证了在患者样本中进行的小鼠研究中发现的G3 MB。我们将开发一个基于云的门户 可视化和探索我们的单细胞和空间数据以及G3 MB的肿瘤-TME相互作用组。二是 建立肿瘤-T细胞相互作用的机制基础和策略，使过继性T细胞治疗能够用于 G3 MB通过发现肿瘤细胞和T细胞中的功能驱动因子和推定靶点。为此我们将 在免疫活性GEMM中应用候选方法和体内CRISPR筛选来识别肿瘤- 将重塑抑制性TME并使G3 MB肿瘤对过继性T细胞敏感的内在调节剂， CAR-T细胞疗法。我们还将测试针对T细胞功能的抑制因子是否能够使 有效的过继性T细胞疗法。我们的研究有望提供新的见解， G3 MB中肿瘤-TME相互作用的机制，并表现出合理的免疫机会。